Rapid Estimation of Tadalafil by Reverse-phase High-performance Liquid Chromatography Method in Bulk and Tablet Formulation

The simple, selective, precise and accurate reverse-phase high-performance liquid chromatography method was developed and validated for analysis of tadalafil in bulk and tablet dosage form. The column was Inertsil C18 (150×4.6 mm; 5 μm) in isocratic mode. The mobile phase used was phosphate buffer (10 mM, pH 3.2) and acetonitrile (50:50% v/v) at the flow rate of 1.0 ml/min with ultraviolet detection at 295 nm at ambient temperature. The retention time for tadalafil was found to be 4.01 min. Linearity was observed in the concentration range from 60 to 140 μg/ml for tadalafil with a correlation coefficient of (r²) 0.9998. The method was validated according to International Conference on Harmonisation guidelines in terms of linearity, accuracy, precision and specificity. Hence, the proposed method can be utilized for routine quality control of tadalafil in bulk and tablet dosage form.     

Ion-Pairing Reverse-Phase High Performance Liquid Chromatography Method for Simultaneous Estimation of Atenolol and Indapamide in Bulk and Combined Dosage Form

A sensitive, accurate, precise and validated ion-pairing reverse-phase liquid chromatographic method for the quantitative determination of atenolol and indapamide in bulk and tablet dosage form was developed. The proposed ion-pairing reverse-phase high performance liquid chromatography method utilises C₁₈ column with 5 μm, 150×4.6 mm i.d. column and mobile phase consisting of 0.1% w/v solution of octane sulphonic acid, sodium salt and methanol (55:45 v/v), (pH 2.8) and ultraviolet detection at 235 nm. A linearity range of 1-250 μg/ml and 1-25 μg/ml for atenolol and indapamide, respectively, was obtained. The mean recoveries are 100.48 and 99.82% for atenolol and indapamide, respectively. The method was validated as per International Conference on Harmonization guidelines.     

Validated Stability-indicating Reverse-phase Ultra-performance Liquid Chromatography Method for Simultaneous Determination of Sodium Methylparaben,Sodium Propylparaben and Ketorolac Tromethamine in Topical Dosage Forms

A sensitive, fast, and stability-indicating isocratic reverse-phase ultra-performance liquid chromatography method was developed and validated for quantitative simultaneous determination of sodium methylparaben, sodium propylparaben and ketorolac tromethamine in topical dosage forms. Separation of all peaks was achieved by using acquity ethylene bridged hybrid C18 (50×2.1 mm, 1.7 μ) as stationary phase, mobile phase used was triethylamine buffer (pH 2.5):tetrahydrofuran:methanol (665:35:300, v/v/v) with isocratic mode at a flow rate of 0.40 ml/min. All component were detected at 252 nm with 10 min run time. The described method was found to be linear in the concentration range of 248-744 μg/ml for ketorolac tromethamine, 20.8-62.4 μg/ml for sodium methylparaben and 2.38-7.13 μg/ml for sodium propylparaben with correlation coefficients more than 0.999. Method was validated in terms of specificity, linearity, accuracy, precision, solution stability, filter equivalency, and robustness as per International Conference on Harmonization guideline. Formulation was exposed to the stress conditions of peroxide, acid, base, thermal, and photolytic degradation and proven all components were well separated in the presence of degradants.     

Rapid and Sensitive Reverse-phase High-performance Liquid Chromatography Method for Estimation of Ketorolac in Pharmaceuticals Using Weighted Regression

A reliable, rapid and sensitive isocratic reverse phase high-performance liquid chromatography method has been developed and validated for assay of ketorolac tromethamine in tablets and ophthalmic dosage forms using diclofenac sodium as an internal standard. An isocratic separation of ketorolac tromethamine was achieved on Oyster BDS (150×4.6 mm i.d., 5 μm particle size) column using mobile phase of methanol:acetonitrile:sodium dihydrogen phosphate (20 mM; pH 5.5) (50:10:40, %v/v) at a flow rate of 1.0 ml/min. The eluents were monitored at 322 nm for ketorolac and at 282 nm for diclofenac sodium with a photodiode array detector. The retention times of ketorolac and diclofenac sodium were found to be 1.9 min and 4.6 min, respectively. Response was a linear function of drug concentration in the range of 0.01-15 μg/ml (R²=0.994; linear regression model using weighing factor 1/x²) with a limit of detection and quantification of 0.002 μg/ml and 0.007 μg/ml, respectively. The % recovery and % relative standard deviation values indicated the method was accurate and precise.     

Development and Validation of RP-HPLC Method for the Simultaneous Estimation of Montelukast Sodium and Ebastine in Tablet Dosage Form

A rapid and sensitive RP-HPLC method with UV detection (244 nm) for routine analysis of montelukast sodium and ebastine in a pharmaceutical formulation (Ebast-M) was developed. Chromatography was performed with mobile phase containing a mixture of methanol:acetonitrile:ammonium acetate (80:10:10, % v/v/v), pH of mobile phase was adjusted 5.5 using glacial acetic acid and flow rate was 1.2 ml/min. The method was validated for linearity, accuracy, robustness and intermediate precision. The linearity was established over the concentration range of 0.01−0.06 mg/ml for both drugs. The correlation coefficients (r²) for ebastine and montelukast were 0.9989 and 0.9955, respectively. Statistical analysis of the data showed that the method was precise, accurate, reproducible and selective for the analysis of ebastine and montelukast drugs. The method was successfully employed for the determination of ebastine and montelukast in commercially available tablet dosage form.     

HPLC法测定胞磷胆碱钠氯化钠注射液中胞磷胆碱钠的含量

目的:建立胞磷胆碱钠氯化钠注射液中胞磷胆碱钠含量测定方法。用于生产及市场监控。方法:选用 C_(15)(250×46mm)5μm 色谱柱,0.02mol/L 磷酸二氢钾水溶液为流动相,检测波长275nm,采用高效液相色谱法测定胞磷胆碱钠的含量;结果:胞磷胆碱钠在28.4～568.0μg/mL 浓度范围内呈良好线性,其回收率为100.07%(RSD=0.399 n=9):结论:上述方法简便、快捷,结果准确,可用于胞磷胆碱钠氯化钠注射液中胞磷胆碱钠含量测定和质量控制。     

Development and Validation of a RP-HPLC Method for Estimation of Montelukast Sodium in Bulk and in Tablet Dosage Form

The present work describes a simple, precise and accurate HPLC method for estimation of montelukast sodium in bulk and in tablet dosage form. The separation was achieved by using octadecylsilane column (C18) and acetonitrile:1 mM sodium acetate adjusted to pH 6.3 with acetic acid in proportion of 90:10 v/v as mobile phase, at a flow rate of 1.5 ml/min. Detection was carried out at 285 nm. The retention time of montelukast sodium was found to be 3.4 min. The limit of detection was found 1.31 µg/ml and limit of quantification 3.97 µg/ml. The accuracy and reliability of the proposed method was ascertained by evaluating various validation parameters like linearity (1-100 µg/ml), precision, accuracy and specificity according to ICH guidelines. The proposed method provides an accurate and precise quality control tool for routine analysis of montelukast sodium in bulk and in tablet dosage form.     

High-Performance Liquid Chromatographic and High-Performance Thin-Layer Chromatographic Method for the Quantitative Estimation of Dolutegravir Sodium in Bulk Drug and Pharmaceutical Dosage Form

Simple, sensitive, precise, and specific high-performance liquid chromategraphic (HPLC) and high-performance thin-layer chromatographic (HPTLC) methods for the determination of dolutegravir sodium in bulk drug and pharmaceutical dosage form were developed and validated. In the HPLC method, analysis of the drug was carried out on the ODS C₁₈ column (150 × 4.6 mm, 5 μm particle size) using a mixture of acetonitrile: water (pH 7.5) in the ratio of 80:20 v/v as the mobile phase at the flow rate 1 mL/min at 260 nm. This method was found to be linear in the concentration range of 5–35 μg/mL. The peak for dolutegravir sodium was observed at 3.0 ± 0.1 minutes. In the HPTLC method, analysis was performed on aluminum-backed plates pre-coated with silica gel G60 F₂₅₄ using methanol: chloroform: formic acid in the proportion of 8:2:0.5 v/v/v as the mobile phase. This solvent system was found to give compact spots for dolutegravir sodium with the Rf value 0.77 ± 0.01. Densitometric analysis of dolutegravir sodium was carried out in the absorbance mode at 265 nm. Linear regression analysis showed good linearity with respect to peak area in the concentration range of 200–900 ng/spot. The methods were validated for precision, limit of detection (LOD), limit of quantitation (LOQ), accuracy, and specificity. Statistical analysis showed that both of the methods are repeatable and specific for the estimation of the said drug. The methods can be used for routine quality control analysis of dolutegravir sodium.     

高效液相色谱法测定注射用头孢噻肟钠舒巴坦钠中2-巯基苯并噻唑的残留量

目的：建立高效液相色谱法测定注射用头孢噻肟钠舒巴坦钠中2-巯基苯并噻唑的残留量。方法：高效液相色谱法的色谱柱：用十八烷基硅烷键合硅胶为填充剂Kromasil 100-5-C18（250 mm × 4.6 mm,5 μm）;流动相：甲醇-10 mmol·L-1乙酸铵（55∶45）;柱温：30 ℃;检测波长：320 nm;流速：1.0 mL·min-1;进样量：10 μL。结果：定量限、检测限、线性关系、耐用性、重复性、进样精密度均良好。结论：本法可用于注射用头孢噻肟钠舒巴坦钠中2-巯基苯并噻唑限度的测定。     

Development and Validation of a Reversed-Phase High-performance Liquid Chromatography Method for Determination of Desmopressin in Chitosan Nanoparticles

A simple isocratic reversed-phase high performance liquid chromatographic method was developed for determination of released desmopressin from chitosan nanoparticles in the in vitro media. The chromatographic separation was achieved with acetonitrile/water (25:75, v/v), in which water contained 0.1% v/v trifluoroacetic acid with pH=2.5 as mobile phase, a Chromolith^® Performance RP-18e column (150×4.6 mm; 5 μm) kept at 40° and ultraviolet detection at 220 nm. The compound was eluted isocritically at a constant flow rate of 1.6 ml/min. The method was validated according to the International Conference on Harmonisation guidelines. The validation characteristics included accuracy, precision, linearity rang, selectivity, limit of detection, limit of quantitation and robustness. The calibration curve was linear (r>0.9999) over the concentration rang 0.5-100 μg/ml. The limit of detection and limit of quantitation in the release media were 0.05 and 0.5 μg/ml, respectively. The proposed method had an accuracy of and intra- and inter-day precision <4.2. Furthermore, to evaluate the performance of the proposed method, it was used in the analysis of desmopressin level in real samples containing chitosan nanoparticles in the in vitro media.     

Reversed-phase High-performance Liquid Chromatography Method for Analysis of Curcuminoids and Curcuminoid-loaded Liposome Formulation

A simple, sensitive, precise and specific method for the determination of curcuminoids and curcuminoid-loaded liposome formulation was developed using reverse-phase high-performance liquid chromatography method. The analysis was performed isocratically on Zorbax Eclipse XDB-C18 column (150×4 mm, 5 μm), analytical column using UV detector and mobile phase consisting of 0.1% orthophosphoric acid and acetonitrile. The proposed method for curcuminoids was validated for linearity in the range from 50 to 300 ΅g/ml with correlation coefficient above 0.997. Intraday and interday precision studies showed the relative standard deviation less than 2%. The limit of detection and limit of quantitation values were 2.5 and 8.25 ΅g/ml, respectively. Forced degradation study for curcuminoids and liposomal curcuminoids sample was carried out and observed that proposed method was also suitable for finding degradation products in the sample. Proposed method was successfully applied to estimate curcuminoids content without any interference of other excipients from liposomal formulation. Therefore, the method developed is well suited for curcuminoids and its liposome estimation.     

Development and Validation of HPTLC Method for the Estimation of Almotriptan Malate in Tablet Dosage Form

A new, simple, precise and accurate high performance thin layer chromatographic method has been proposed for the determination of almotriptan malate in a tablet dosage form. The drug was separated on aluminum plates precoated with silica gel 60 GF(254) with butanol:acetic acid:water (3:1:1) was used as mobilephase. Quantitative analysis was performed by densitometric scanning at 300 nm. The method was validated for linearity, accuracy, precision and robustness. The calibration plot was linear over the range of 100-700 ng/band for almotriptan malate. The method was successfully applied to the analysis of drug in a pharmaceutical dosage form.     

Development and Validation of a HPTLC Method for Simultaneous Quantitation of Flunarizine Dihydrochloride and Propranolol Hydrochloride in Capsule Dosage Form

A simple, precise, accurate, and rapid high-performance thin layer chromatographic method has been developed and validated for the simultaneous quantitation of flunarizine dihydrochloride and propranolol hydrochloride in a combined capsule dosage form. The method was carried out on precoated silica gel 60 F₂₅₄ TLC aluminum plate, (20×10 cm²). The solvent system was ethyl acetate:methanol:glacial acetic acid in the proportion of 8:1:1, (v/v/v). R_f value for flunarizine dihydrochloride and propranolol hydrochloride was found to be 0.62±0.02 and 0.18±0.02, respectively. The linearity regression analysis for calibration showed 0.999 and 0.999 for flunarizine dihydrochloride and propranolol hydrochloride with respect to peak area and height in the concentration range of 50-350 ng/spot and 500-3500 ng/spot, respectively. Accuracy of recovery studies was found to be 98-100.28 and 99.11-99.45% for flunarizine dihydrochloride and propranolol hydrochloride, respectively. The amounts of drug in marketed formulation were 100.5 and 101.25% of flunarizine dihydrochloride and propranolol hydrochloride, respectively. The method developed can be used for routine analysis in bulk drug and capsule dosage form.     

Development and Validation of Stability-indicating High Performance Liquid Chromatographic Method for the Estimation of Everolimus in Tablets

The present study depicts the development of a validated reversed-phase high performance liquid chromatographic method for the determination of the everolimus in presence of degradation products or pharmaceutical excipients. Stress study was performed on everolimus and it was found that it degrade sufficiently in oxidizing and acidic conditions but less degradation was found in alkaline, neutral, thermal and photolytic conditions. The separation was carried out on Hypersil BDS C18 column (100×4.6 mm, 5 μ) column having particle size 5 μ using acetate buffer:acetonitrile (50:50 v/v) with pH 6.5 adjusted with orthophosphoric acid as mobile phase at flow rate of 1 ml/min. The wavelength of the detection was 280 nm. A retention time (R_t) nearly 3.110 min was observed. The calibration curve for everolimus was linear (r²=0.999) from range of 25-150 μg/ml with limit of detection and limit of quantification of 0.036 μg/ml and 0.109 μg/ml, respectively. Analytical validation parameters such as selectivity, specificity, linearity, accuracy and precision were evaluated and relative standard deviation value for all the key parameters were less than 2.0%. The recovery of the drug after standard addition was found to be 100.55%. Thus, the developed RP-HPLC method was found to be suitable for the determination of everolimus in tablets containing various excipients.     

高效凝胶色谱法测定注射用头孢哌酮钠他唑巴坦钠中的高聚物

目的建立测定注射用头孢哌酮钠他唑巴坦钠(4∶1)中高聚物的高效凝胶色谱法。方法采用Ohpak SB-802.5HQ凝胶色谱柱,以pH=7.0的磷酸盐缓冲液[0.03 mol/L磷酸氢二钠溶液-0.03 mol/L磷酸二氢钠溶液(61∶39)]-乙腈(70∶30)为流动相,主成分自身对照法定量。结果建立了在该系统下以保留时间小于头孢哌酮钠所有杂质峰面积的总和进行质量控制的检测方法。结论所用方法简便,结果可靠,可用于注射用头孢哌酮钠他唑巴坦钠(4∶1)中高聚物的检测。     

Development and Validation of RP-HPLC Method for the Determination of Methamphetamine and Propranolol in Tablet Dosage Form

A new isocratic reversed-phase HPLC method with diode-array UV detection was developed and validated for the determination of methamphetamine and propranolol in tablet dosage forms. Chromatography was carried out on an XTerra RP18 (150×4.6 mm, 5 μm) column using 50 mM pyrrolidine (pH 11.5) - acetonitrile (50:50, v/v) as mobile phase at a flow rate of 1 ml/min. Spectrophotometric detection was performed at a wavelength of 214 nm. The linearity was established over the concentration range of 0.075-0.60 mg/ml for both drugs. The correlation coefficients (r(2)) were ≥0.9998 in each case. The relative standard deviation values for intermediate precision studies were <1%. Statistical analysis of the data showed that the method was precise, accurate, reproducible and selective for the analysis of methamphetamine and propranolol drugs. The method was successfully employed for the determination of propranolol and methamphetamine in commercially available tablet dosage form.     

高效液相色谱法测定大白鼠肠液中乳梨醇浓度

目的：建立高效液相色谱法测定大白鼠小肠液和盲肠液中乳梨醇浓度。方法：采用ＷａｔｅｒｓＳｕｇａｒ－Ｐａｋ１色谱柱,示差折光检测器,以水为流动相,柱温９０℃,流速０５ｍＬ·ｍｉｎ－１。在００２～２４ｍｇ·ｍＬ－１浓度范围内线性良好,最低检测浓度为２０μｇ·ｍＬ－１,回收率为９７５％（ＲＳＤ＝０２７％,ｎ＝５）。结果：乳梨醇在小肠内经０ｈ和４ｈ时的浓度分别为（１１４±０１５）ｍｇｍＬ－１和（０９７±０１５）ｍｇ·ｍＬ－１,在盲肠内经０ｈ和４ｈ时的浓度分别为（０９５±０２６）ｍｇ·ｍＬ－１和（０３０±００８）ｍｇ·ｍＬ－１。结论：肠道给药４ｈ后乳梨醇在小肠内的浓度基本不变,在盲肠内下降了１／３,可能在大肠杆菌中有关酶的作用下降解吸收。     

高效液相色谱法同时测定肺力咳合剂中苯甲酸钠等6种防腐剂含量

目的：建立高效液相色谱（HPLC）同时测定肺力咳合剂中苯甲酸钠等6种防腐剂含量的方法。方法：采用HPLC,色谱柱为Agilent XDB-C18色谱柱（250 mm×4.6 mm,5 μm）,流动相：甲醇-0.025 mol/L磷酸二氢钠溶液（梯度洗脱）;流速：1.0 mL/min;柱温30 ℃,检测波长228 nm、255 nm。结果：肺力咳合剂中苯甲酸钠在0.063 54～211.800 00 μg（r=0.999 9）、山梨酸钾在0.066 06～220.200 00 μg（r=0.999 9）、对羟基苯甲酸甲酯在0.062 70～209.000 00 μg（r=0.999 9）、对羟基苯甲酸乙酯在0.066 12～220.400 00 μg（r=0.999 9）、对羟基苯甲酸丙酯在0.067 98～226.600 00 μg（r=0.999 9）,对羟基苯甲酸丁酯在0.063 30～211.000 00 μg（r=0.999 9）范围内线性良好,平均回收率分别为100.72%（RSD=0.82%）、104.82%（RSD=1.59%）、101.11%（RSD=0.25%）、102.39%（RSD=1.33%）、102.56%（RSD=1.35%）、101.09%（RSD=0.34%）。10批样品中均未检出山梨酸钾、对羟基苯甲酸甲酯、对羟基苯甲酸丁酯,苯甲酸钠、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯的含量分别为2.132 6~2.602 6 mg/mL、0.110 8~0.126 0 mg/mL、0.399 9~0.408 9 mg/mL。结论：该方法具有操作简单,专属性强,准确度高的优点,可以更全面的控制肺力咳合剂中防腐剂的含量。     

Stability Indicating RP-HPLC Method for Simultaneous Determination of Simvastatin and Ezetimibe from Tablet Dosage Form

A simple, specific and sensitive reverse phase high performance liquid chromatographic method was developed and validated for simultaneous determination of ezetimibe and simvastatin from pharmaceutical dosage forms. The method uses C18 ODS Hypersil column and isocratic elution. The mobile phase composed of acetonitrile:phosphate buffer (pH 4.5, 0.01M) in the ratio of 65:35 v/v was used at a flow rate of 1.0 ml /min. UV detector was programmed at 232 nm for first 10 min and at 238 nm for 10 to 20 min. All the validation parameters were in acceptable range. The developed method was effectively applied to quantitate amount of ezetimibe and simvastatin from tablets. The method was also applied suitably for determining the degradation products of ezetimibe and simvastatin.     

Validation of a Stability-Indicating RP-HPLC Method for the Simultaneous Determination of Trimethoprim and Sulfadimethoxine Sodium in Oral Liquid Dosage Form

A simple, specific, accurate, and stability-indicating RP-HPLC method was developed and validated for the simultaneous determination of Trimethoprim (TMP) and Sulfadimethoxine sodium (SDMS) in Vetricine^® oral solution product. The desired separation was achieved on an ODS column (250×4.6 mm i.d., 5 μm) at room temperature. The optimized mobile phase consisted of an isocratic solvent mixture of water:acetonitrile:triethylamine (700:299:1, v/v/v), adjusted to a pH of 5.7 ± 0.05 with 0.2N acetic acid. The mobile phase was fixed at 0.8 ml/min and the analytes were monitored at 254 nm using a photodiode array detector. The effects of the chromatographic conditions on the peaks USP tailing factor, column efficiency, and resolution were systematically optimized. Forced degradation experiments were carried out by exposing TMP, SDMS standards, and the oral solution formulation to thermal, photolytic, oxidative, and acid-base hydrolytic stress conditions. The degradation products were well-resolved from the main peaks and the excipients, thus proving the reliable stability-indicating method. The method was validated as per ICH and USP guidelines (USP34/NF29) and found to be adequate for the routine quantitative estimation of TMP and SDMS in commercially available Vetricine® oral liquid dosage form.